Cullin1蛋白在胃癌中的表达及临床意义

目的:研究Cullin1在胃癌细胞及胃癌组织中的表达,并分析Cullin1、临床分期与患者生存率之间的相关性.方法:采用Western blot法检测SGC-7901、BGC-823胃癌细胞与正常胃黏膜上皮细胞GES-1中、胃癌组织与癌旁组织中Cullin1蛋白表达差异.我们利用已经构建的胃癌数据库,采用免疫组织化学法检测792例胃癌组织中Cullin1表达.结果:我们研究发现,SGC-7901、BGC-823胃癌细胞中Cullin1表达水平均高于胃上皮细胞株GES-1(P<0.01).胃癌组织中Cullin1蛋白表达水平均高于癌旁组织(P<0.01).Cullin1过表达与胃癌TNM分期(P=0.011)、浸润深度(P=0.035,T1-T3与T4)及淋巴结转移(P=0.036)显著相关.此外,我们发现Cullin1的高表达与胃癌患者较差的总生存时间及3年生存率明显相关(P=0.042,0.026).Cox回归分析显示,Cullin1表达是胃癌患者3年生存率的一个独立的预后因子(P=0.028).结论:我们的数据表明,Cullin1可作为胃癌淋巴结转移、预后以及潜在治疗的一个重要标志和研究目标.

Expression and location of α-fetoprotein during rat colon development

AIM： To investigate the expression of α-fetoprotein （AFP）, a cancer-associated fetal glycoprotein, and its involvement during rat colon development. METHODS： Colons from Sprague-Dawley rat fetuses, young and adult （8 wk old） animals were used in this study. Expression levels of AFP in colons of different development stage were detected by reversetranscriptase PCR （RT-PCR） and Western blotting. To identify the cell location of AFP in the developing rat colons, double-immunofluorescent staining was performed using antibodies to specific cell markers and AFP, respectively. RESULTS： The highest levels of AFP mRNA were detected in colons of rats at embryonic day 18.5 （e18.5）. Compared to e18.5 d, the AFP expression was significantly decreased during rat development （85% for e20.5, P 〈 0.05, 58% for postnatal day 0.5 （P〈0.5）, P 〈 0.05, 37% for P7, P 〈 0.05, 24% for P14, P 〈 0.05, and 11% for P21, P 〈 0.05） and undetected in adult rats. Only the 72-kDa isoform of AFP was detected by Western blotting, the expression pattern was similar to AFP mRNA and conformed to the results of mRNA expression. The AFP positive staining was identical to different distribution patterns in fetuses, young and adult animals and positive staining for both AFP and vimentin was overlapped in mesenchymal cells at each stage tested. CONCLUSION： This study has for the first time demonstrated that AFP is localized in the mesenchyme of rat colon from the embryo to the weaning stage by immunofluorescence and presents 72-kDa isoform in the developing rat colons by Western blotting. The dynamic expression of AFP in the various developmental stages of the colon indicates that AFP might be involved in many aspects of colon development.

Bone Marrow Urokinase Plasminogen Activator Receptor Levels are Associated with the Progress of Multiple Myeloma

Objective To determine the mRNA and protein levels of urokinase plasminogen activator receptors(u PAR) in bone marrow fluid and bone marrow tissue from multiple myeloma(MM) patients and assess association of u PAR level with prognosis of MM.Methods u PAR levels in bone marrow fluid of 22 MM patients at the stable and progressive stages and 18 iron deficiency anemia patients with normal bone marrow(control) were examined by ELISA.Furthermore,u PAR expression in bone marrow tissue was investigated by RT-PCR and Western blot,respectively.The distribution of u PAR in MM cells was examined using immunofluorescence staining.The pathological changes in different stages of MM patients were studied by HE staining.Results u PAR level in bone marrow fluid of MM patients(1.52±0.32 μg/ml) was found to be higher than that in the control group(0.98±0.15 μg/ml).Interestingly,u PAR protein(0.686±0.075 vs.0.372±0.043,P<0.05) and m RNA(2.51±0.46 vs.4.46±1.15,P<0.05) expression levels of MM patients at the progressive stage were significantly higher than those at the stable stage.The expression of u PAR in MM bone marrow was confirmed by immunofluorescence staining.Moreover,HE staining revealed a great increased number of nucleated cells and severe impairment of hematopoietic function in the bone marrow of patients with progressive-stage myeloma.Conclusion Our study reveals that u PAR expression is positively correlated with the development and progress of MM.

Expression and role of PAK6 after spinal cord injury in adult rat

Objective： To observe p21-activated kinase 6 （PAK6） expression and its possible role after spinal cord injury （SCI） in adult rat.Methods： Sprague-Dawley rats were subjected to spinal cord injury. To explore the pathological and physiological significance of PAK6, the expression patterns and distribution of PAK6 were observed by Western blot, immunohistochemistry and immunofluorescence.Results： Western blot analysis showed PAK6 protein level was significantly up-regulated on day 2 and day 4,then reduced and had no up-regulation till day 14. Immunohistochemistry analysis showed that the expression of PAK6 was significantly increased on day 4 compared with the control group. Besides, double immunofluorescence staining showed PAK6 was primarily expressed in the neurons and astrocytes in the control group. While after injury, the expression of PAK6 was increased significantly in the astrocytes and neurons, and the astrocytes were largely proliferated. We also examined the expression of proliferating cell nuclear antigen （PCNA） and found its change was correlated with the expression of PAK6. Importantly, double immunofluorescence staining revealed that cell proliferation evaluated by PCNA appeared in many PAK6-expressing cells on day 4 after injury.Conclusion： The up-regulation of PAK6 in the injured spinal cord may be associated with glial proliferation.